Biologie du Développement et Reproduction, UMR1198, INRA ENVA, 78352, Jouy-en-Josas, France.
Transgenic Res. 2010 Oct;19(5):923-31. doi: 10.1007/s11248-010-9368-7. Epub 2010 Jan 27.
This paper reports our attempts to characterize transgene integration sites in transgenic mouse lines generated by the microinjection of large (from 30 to 145 kb) pig DNA fragments encompassing a mammary specific gene, the whey acidic protein gene (WAP). Among the various methods used, the thermal asymmetric interlaced (TAIL-) PCR method allowed us (1) to analyze transgene/genomic borders and internal concatamer junctions for eleven transgenic lines, (2) to obtain sequence information for seven borders, (3) to place three transgenes in the mouse genome, and (4) to obtain sequence data for seven transgene junctions in concatamers. Finally, we characterized various rearrangements in the borders and the inner parts of the transgene. The possibility of such complex rearrangements should be carefully considered when transgenic animals are produced with large genomic DNA fragments.
本文报告了我们通过微注射包含乳特异性基因乳白蛋白蛋白基因（WAP）的大型（30 至 145kb）猪 DNA 片段，在转基因小鼠系中对转基因整合位点进行特征分析的尝试。在使用的各种方法中，热不对称交错（TAIL-）PCR 方法使我们能够：(1) 分析十一条转基因线的转基因/基因组边界和内部串联连接；(2) 获得七个边界的序列信息；(3) 将三个转基因放置在小鼠基因组中；(4) 获得七个串联连接的转基因连接的序列数据。最后，我们对边界和转基因内部的各种重排进行了特征分析。在使用大的基因组 DNA 片段生产转基因动物时，应该仔细考虑这种复杂重排的可能性。
